Mechanism of lentivirus mediated flot1 gene expression on invasion and apoptosis of gastric cancer cells
- Author:
You-Jie FAN
1
Author Information
- Publication Type:Journal Article
- Keywords: Apoptosis; Epithelial mesenchyme transformation; FLOT1 gene; Gastric cancer; Invasion
- From: Chinese Pharmaceutical Journal 2019;54(16):1299-1304
- CountryChina
- Language:Chinese
- Abstract: OBJECTIVE: To investigate inhibitory effect of FLOT1 gene expression on invasion and apoptosis of gastric cancer cells. METHODS: The designed FLOT1 siRNA lentiviral vector (si-FLOT1 group) was transfected into human gastric cancer MKN-45 cells, at the same time, the negative control lentivirus vector (negative group) was transfected, and the blank group was set up. Western blotting was used to detect the expression of FLOT1, E-cadherin, α-SMA, cleaved caspase 3, Bcl-2 and Bax proteins. After cells were transfected for 24, 48, 72 and 96 h, cell viability was detected by CCK-8 assay. After cells were transfected for 48 h, transwell chamber, flow cytometry and DCFH-DA assay were used to detect the invasiveness, apoptosis rate and ROS content, respectively. RESULTS: FLOT1 siRNA lentiviral vector inhibited significantly the expression of FLOT1 in MKN-45 cells, which was significantly different from the blank group (P<0.05). Compared with si-FLOT1 group and the blank group, the cell vitality decreased, the invasion ability decreased, the apoptosis rate increased, the expression of E-cadherin, cleaved caspase 3 and Bax protein increased, the expression of α-SMA and Bcl-2 protein decreased, and the content of ROS increased, and the difference was statistically significant (P<0.05). CONCLUSION: Down regulation of FLOT1 gene expression can reduce the invasiveness of gastric cancer cells by inhibiting EMT, and promote apoptosis by regulating the expression of apoptosis related proteins and increasing the ROS content of cells.
